when sample containing sulfated Glycosaminoglycans is mixed with the DMMB dye it forms an insoluble complex which precipitates immediately, hindering spectrophotometric measurement.
How to Overcome this?
Dear Sir,
please note that that DMMB (dimethylmethylene blue ) is sold as zinc chloride double salt (p.e. Sigma). Are you mixing the dye salt and sulfated glycosaminoglycans in WATER?
In this case, you ´ll always obtain a precipitate in water. It is due to the dimethyl groups of the dye and the interaction of DMMB as cationic dye with the sulfated glycosaminoglycans (see the solubility of this compound in water alone). The presence of zinc double salt (if you use it instead chloride) salts out the precipitate too.
Please give Valeria Scagliotti in R.G. search for researchers… and for 2013 you will obtain a lot of hits about DMMB and glycosaminoglycans assay .
RG and previous answers of researchers can help you for this problem.
Are you working with cuvettes or microplates?
JRG
Thank you very much for your support. I’m going to work with microplates..
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